Question

The PCR reaction for Lab 2 is listed below. For each component besides water, briefly describe the component’s role in the PCR reaction. PCR amplification recipe (stock concentrations):
14.4 μL high quality nuclease‐free water
6.0 μL 5x Phusion HF buffer
3.0 μL dNTPs (2.5 mM each of four dNTPs – dATP, dCTP, dGTP, dTTP)
0.3 μL Phusion DNA polymerase
0.3 μL pCD122 (a plasmid that serves as template DNA)
3.0 μL sense primer CD474 (5 μM)
3.0 μL antisense primer CD475 (5 μM)
30.0 μL total reaction volume

Answer 1

Answer:

PCR is known as polymerase chain reaction used to amplify DNA sequence of our interest into multiple copies. This technique is been commonly used by researchers to study in depth about the gene of interest during their research work.

Explanation:

PCR:- It is known as polymerase chain reaction, used to amplify DNA sequence of our interest into multiple copies mainlty to millions or trillions

Components

Phusion HF buffer:- This buffer Create optimal reaction conditions for high fidelity amplification of DNA

dNTPs:- They are the building blocks in the synthesis of new copies of DNA.There are four dNTPs used in the amplification process that is dATP, dGTP, dCTP, dTTP. these building blocks are added in equal proportion during the PCR reaction

Phusion DNA polymerase:- This is the enzyme used in DNA amplification, it is generally a fusion of  DNA binding domain to a portion of pyrococcus like proof reading polymerase. This enzyme is tolerant to various inhibitors, allowing strong amplification of DNA of interest with minimal optimization

pCD122:- Plasmid that serves as a template DNA for the amplification of desired DNA sequence of our interest.

sense primer CD474:- Sense primer is also known a reverse primer, it attaches to the stop codon of the complementary strand of DNA

antisense primer CD475:- This primer is also known as forward primer, it attaches to the start codon of the template DNA

All these components is added in such a way that the total mixture should have a reaction volume of 30.0μl